JCS033316 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Silencing of BDNF protein in neuroblastoma cell lines. Neuroblastoma cell lines SH-SY-5Y and SK-N-BE were transfected with the construct pEGFP-N1-BDNFcds encoding BDNF in fusion with GFP. In the same conditions, cells were cotransfected with the siRNA specific for BDNF or for GAPDH mRNA. (A) Cells transfected with pEGFP-N1-BDNFcds. (B) Cells cotransfected with pEGFP-N1-BDNFcds and the siRNA against BDNF CDS mRNA. (C) Cell cotransfected with pEGFP-N1-BDNFcds and the siRNA against GAPDH mRNA. (D) Control cultures that received the transfection medium without plasmid. (E) Quantification of the viability assay was performed by counting positive cells with respect to total number of cells determined by counting nuclei labelled with Hoechst 33258 (Each column is the mean ± s.e.m. of three cultures). Statistical analysis was performed with ANOVA (*P<0.05; **P<0.01).

• Supplemental Figure S2 -

  Fig. S2. Viability assay, using the MTT quantification method, of SH-SY-5Y cells treated with siRNAs against each single BDNF mRNA isoforms at 6 hours in (A) control medium (10% FCS), (B) control medium with CIS, (C) serum-free medium (SFM) but no drugs, (D) SFM with CIS. Each column represents the mean ± s.e.m. of three cultures. Statistical analysis was performed with ANOVA (*P<0.05; **P<0.01).